【作者】 周勇；

【导师】 曾海涛；

【作者基本信息】 中南大学 ， 生物化学与分子生物学， 2012， 硕士

【摘要】 镉是一种对环境和人体有害的重金属,在有机体内生物半衰期长达10～30年,是极易在生物体内储积的环境污染物之一,可引起机体多组织多器官的广泛损伤,尤其是对男性生殖系统损伤最为敏感。目的：(1)观察镉体外对小鼠精子获能和项体反应的影响；(2)研究镉体外对小鼠精子获能影响是否通过蛋白丝氨酸磷酸化途径；(3)小鼠体内染镉对精子获能及DNA损伤影响。方法：(1)分别在镉离子终浓度为0、10、50、500、2500和5000μmol/L时体外培养小鼠精子获能80min,用A23187诱导顶体反应10mmin,考马斯亮蓝染色后观测小鼠精子发生获能的比例；(2)小鼠精子在体外培养获能80min后,加入镉离子终浓度分别为0、50、500和2500μmol/L,再用A23187诱导顶体反应1Omin,考马斯亮蓝染色后观测小鼠精子发生顶体反应的比例；(3)用0、10、50、500和5000μmol/L镉离子浓度处理小鼠精子获能90min后,用Western blot法确定发生丝氨酸磷酸化蛋白的差异；(4)人工水质染镉6周后检测小鼠精子获能比例及DNA损伤指数。结果：(1)所选镉离子浓度10、50、500、2500和5000μmol/L对小鼠精子发生获能有显著影响(P<0.05)；(2)所选镉离子浓度50、500和2500μmol/L对小鼠精子顶体反应发生无显著影响(P>0.05)；(3)发现随着镉离子浓度升高14kDa,90kDa处蛋白丝氨酸磷酸化增强。(4)慢性染镉后获能受抑制,DNA损伤指数升高。结论：(1)镉在体外对小鼠精子发生顶体反应没有抑制作用；(2)镉在体外对小鼠精子获能有抑制作用且可能是通过蛋白丝氨酸磷酸化途径；(3)慢性染镉会抑制小鼠精子获能并会损伤精子DNA。更多还原

【Abstract】 Cadmium is a heavy metal harmful to the environment and human, Its half-life is up to10-30years in vivo, and is one of the accumulation of environmental pollutants.It can cause extensive body damage, especially the male reproductive system.Objective:(1)To study the effects of cadmium on acrosome reaction and capacitation in vitro in mouse spermatozoa;(2)To study whether cadmium affect proteins serine phosphorylation pathway in the sperm capacitation;(3)To study the effects of cadmium on capacitation and DFI in vivo in mouse sperm.Methods:(1) Mouse cauda epididymal spermatozoa were suspended in EKRB medium supplemented with3mg BSA/mL in the presence of0、10、50、500、2500and5000μmol/L cadmium respectively and incubated at37℃for80min, and stimulated with A23187for10min. Acrosome reaction of the sperm was analyzed by Coomassie blue staining;(2) Cauda epididymal spermatozoa were suspended in EKRB medium supplemented with3mg BSA/mL and incubated at37℃for80min, then be treated with cadmium at the concentration of0、10、50、500、 and2500μmol/L respectively and stimulated with A23187for10min. Acrosome reaction of the sperm was analyzed by Coomassie blue staining;(3) Cauda epididymal spermatozoa were capacitated in vitro for90min with cadmium at the concentration of0、10、50、500、5000μmol/L then serine phosphorylation of sperm components was detected by Western blot after SDS-PAGE;(4) Sperm capacitation and DFI were observed after mouse drank water containg cadmium for6weeks.Results:(1)Cadmium significantly inhibited mouse sperm capacitation at the concentration of10、50、500、250、5000μmol/L (P<0.05);(2)Cadmium exhibited no significant influence on acrosome reaction at the concentration of50、500、2500μmol/L(P>0.05);(3)Serine phosphorylation of14kDa,90kDa proteins increased with cadmium at the concentration of0、10、50、500、5000μmol/L;(4) Cadmium significantly inhibited sperm capacitation and damaged the sperm DNA in vivo(P<0.05).Conclusions:(1)Cadmium has no direct effect on mouse sperm acrosome reaction;(2)Cadmium may be affect mouse sperm capacitation by proteins serine phosphorylation in vitro;(3) Cadmium can inhibit mouse sperm capacitation and damage sperm DNA in vivo.更多还原